This Project is targeted to advanced engineering research in the areas of microfabrication and molecular biology analysis. The primary goal of is to develop components that will advance the state-of-the-art of microfabricated biochemical analysis. The Specific Aims are: Aim 1. Development of advanced fabrication methods compatible with integrated biochemical systems. Bonding of glass and silicon surfaces to generate improved hermetically sealed channels will be developed in this aim. The project will also target development of improved thermal isolation methods of regions on the device using techniques such as substrate modification. Aim 2. Development of advanced components for use in integrated systems. Advanced microfluidic control including valves, hydrophobic/hydrophilic surfaces, and drop propulsion methods will be tested. Storage of enzymes and other reagents on chips will be investigated for the construction of complete, "off-the-shelf" devices. We will target a 10- to 100-fold improvement in the signal-to-background ratio of integral optical detectors by using microfabricated reflecting surfaces and integral waveguides. We will also continue to improve the resolution and functionality of our electrophoresis section. Aim 3. Development of advanced analytical techniques for microscale systems. Macroscale biochemistry methods--such as single nucleotide primer extension --will be adapted to microscale system formats. In parallel, biochemical methods that take advantage of the novel characteristics of microscale systems will be developed, such as quantitative PCR using constrained geometries.